How do I get started?
Before you make an appointment for training, do some background reading to familiarize yourself with the principles of the technique and determine whether it's likely that it will work for your system. We recommend ForteBio's Application Note 14 (Biomolecular Binding Kinetics Assays on the Octet Platform) for kinetics studies and Applications Brief - Octet Quantitation for quantitation. Both of these are available on the BIF website as well as on the ForteBio website.
How do I arrange for a training session for the instrument?
Contact Debby Pheasant (firstname.lastname@example.org or 617-452-2051) to find out how long training will take and to make an appointment when you and she and the instrument are all free at the same time.
How do I make reservations for the instrument?
The reservation for the initial training session will be made by Debby. After you're trained, you'll make reservations online through the Exchange calendar if you're an MIT person. People from outside MIT will need to ask Debby to make reservations for them.
Are there any restrictions on what materials I can bring into the BIF?
The BIF is a BL1 facility so no prions or other BL2 materials are allowed.
What do I need to bring with me?
You'll need 2 black 96-well plates, Greiner Cat. No. 655209, VWR Cat. No. 82050-784, Fisher Cat. No. 50-320-828. The Octet was set up for these plates specifically. We have some on hand to get you started.
We have on hand the following types of sensor tips: Streptavidin (SA), Super Streptavidin (SSA) and trial packs of Ni-NTA (NTA), Anti-HIS (HIS2), Protein A, Anti-FLAG (FLG), and Anti-GST (GST). If you need another type of tip, you'll need to purchase those yourself.
How do I prepare the samples?
Be sure for kinetics work that all your samples are in the same buffer and that at least one of your binding partners is super-clean. For quantitation studies, be sure the standards are diluted in the same buffer matrix as the unknown. You'll need 200 ul of sample per well. A good general sample buffer is 1X PBS, 1% BSA, 0.05% Tween-20 to reduce non-specific binding.
What are good concentration ranges for samples?
For loading tips, 1-10 ug/ml is a good range. As high as 20 ug/ml (100 nM) will work, also. For association, 10-400 nM is a good range. If you know the KD of your system, start at a 10X concentration and reduce it step-wise. See Applications Note 14 for further details.